Bilateral NSF/BIO-BBSRC: Unravelling the Grass Leaf

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Through a combination of experimental and modelling approaches we aim to produce a mechanistic model for how the key developmental transitions underlying the monocot strategy are generated and genetically controlled. Hypotheses for key developmental and shape transitions will be tested by visualising maize PIN auxin transporter proteins, which act both as markers for tissue cell polarity and as readouts for auxin-based polarity coordination mechanisms. Further tests will be carried out by following the expression of genes known to affect PIN1 polarity and key developmental switches in maize. We will also introduce Cre-Lox reporters to enable induction and visualisation of clonal markers at various stages, allowing hypotheses about growth patterns to be tested. Both Confocal and Optical Projection Tomography imaging will be used to obtain information from the cellular to whole organ scales. We will apply the above methods to wild type and mutants which affect key transitions and determine whether the results confirm or refute particular hypotheses. We will also use the Cre-Lox system to generate timed clonal sectors for genes such as KN1 and analyse their consequences on polarity and growth. In parallel, computational methods will be developed to extract salient features and parameters from images generated by immunolocalisation, GFP markers, clonal analysis and live tissue tracking. The results will provide quantitative measures for hypothesis testing. In conjunction with the experiments described above, hypotheses for key developmental transitions will be further formulated and modified using the GPT computational framework, which allows tissues to be modelled as 2D sheets that deform in 3D. This framework will also be elaborated to allow cellular level models for polarity coordination to be explored, and models for the formation of volumetric 3D outgrowths, such as ligules, to be incorporated.

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